1. Fig. 1. Adhiron coding region and phagemid vector
Fig. 1. Adhiron coding region and phagemid vector. (A) Codon optimized coding sequence and amino acid sequence of the Adhiron92 scaffold with secondary structure elements indicated. The residues that are replaced by the nine randomized amino acids (X) to form LOOP1 and LOOP2 in the Adhiron library are boxed. In place of the N-terminal residues AlaThrGly, the original consensus sequence contained the N-terminal sequence AlaAlaLeuLeuGlyGly. (B) pBSTG1 phagemid vector containing the coding region for Adhiron92 indicating relevant features of the construct.
From: Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications
Protein Eng Des Sel. 2014;27(5): doi: /protein/gzu007
Protein Eng Des Sel | © The Author Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

2. Fig. 5. Characterisation of yeast SUMO-binding Adhirons, Ad-ySUMO 10, 15, 20 and 22. (A) Biotinylated Ad-ySUMOs were used to detect ySUMO (black). Human SUMO 1 (light grey) and human SUMO 2 (dark grey) by ELISA with TMB product detected at 560 nm. (B) Western blots using biotinylated Ad-ySUMO clones against yeast SUMO alone (upper panel) and mixed with 20 µg of HEK293 cell lysate (lower panel). (C) Western blot analysis using biotinylated Ad-ySUMO clones against yeast and human SUMOs 1 and 2. (D) Isothermal calorimetry of Ad-ySUMOs binding to yeast SUMO with the isotherms and the data fits.
From: Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications
Protein Eng Des Sel. 2014;27(5): doi: /protein/gzu007
Protein Eng Des Sel | © The Author Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

3. Fig. 4. Isolation and purification of yeast SUMO-binding Adhirons
Fig. 4. Isolation and purification of yeast SUMO-binding Adhirons. (A) Phage ELISA of Adhirons from 24 clones incubated in wells containing ySUMO (black) or control (grey) showing the TMB product absorbance at 560 nm after 3 min. incubation. (B) Ad-ySUMOs were expressed in BL21 (DE3) cells and cell lysates were heated to 20, 50, 60, 70, 80, 90 and 100°C for 20 min then 5 µl of cleared lysates was separated by 15% SDS–PAGE and Coomasie stained. (C) Ad-ySUMO purification by Ni-NTA beads and analysis of the purified Ad-ySUMOs by 15% SDS–PAGE with Coomassie staining. (D) DSC of Ad-ySUMO clones 10, 15, 20 and 22 together with the Adhiron scaffold.
From: Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications
Protein Eng Des Sel. 2014;27(5): doi: /protein/gzu007
Protein Eng Des Sel | © The Author Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

4. Fig. 3. X-ray crystal structure of Adhiron92 scaffold (PDB ID no
Fig. 3. X-ray crystal structure of Adhiron92 scaffold (PDB ID no. 4N6T) at 1.75 Å resolution. The single alpha helix and the four anti-parallel β strands are shown in white with the insertion sites for library production shown in black. Residues 1–10 and 90–92 are not visible in the structure and are presumably disordered. The structure of Adhiron81 at 2.25 Å resolution (PDB ID no. 4N6U) is essentially identical.
From: Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications
Protein Eng Des Sel. 2014;27(5): doi: /protein/gzu007
Protein Eng Des Sel | © The Author Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

5. Fig. 2. Characterisation of the Adhiron scaffold and library
Fig. 2. Characterisation of the Adhiron scaffold and library. (A) DSC to determine the melting temperature of the Adhiron scaffold (T_m 101°C). (B) Ninety-six random clones were isolated and sequenced both as the phagemid transformed and Adhiron phage library infected E. coli ER2738 cells. The percentage of each amino acid within the variable regions is shown. An ideal library would contain 5.26 ± 2.3% of each amino acid; cysteine was not included in the library. (C) CD analysis of the Adhiron scaffold and of three Adhiron proteins containing inserts from the library. All show high β structure content.
From: Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications
Protein Eng Des Sel. 2014;27(5): doi: /protein/gzu007
Protein Eng Des Sel | © The Author Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

6. Fig. 6. Phage ELISA results for Adhirons identified in screens against (A) growth factor protein FGF1, (B) cell surface receptor CD31 and (C) a peptide. Graphs show absorbance readings of each well after the addition of TMB. Wells containing the target molecule are shown in black and control wells are shown in grey.
From: Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications
Protein Eng Des Sel. 2014;27(5): doi: /protein/gzu007
Protein Eng Des Sel | © The Author Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.